Isolation and Characterization of Adipose derived Mesenchymal Stem cells (ADMSCs) in Madras Red Sheep (Ovis aries).

Abstract

ADMSCs were cultured upto passage 2 (P 2 ). Characterization was done by using transcription factors namely Oct4 and Sox2 at P 0 level and was demonstrated by immunofluroscence technique. Immunophenotyping of ovine ADMSCs was done by using CD34 and CD44 monoclonal antibodies at P 0 and P 1 levels by using flow cytometry. At P0 level, 30.14 per cent ADMSCs were positive to CD44 and 13.83 per cent were positive for CD34 antibodies. At P 1 level, 40 per cent and 7.13 per cent cells were positive for CD44 and CD34 respectively. To conclude, Ovine ADMSCs were identified by Oct4 and Sox2 at P 0 level and CD44 positive cells were more at P 0 and P 1 level.

Isolation of Feline Adipose Tissue Derived Mesenchymal Stem Cells

“…On day five post incubation the cells attained about 60-70 per cent confluency at P1. According Beaulah et al, (2016) in Madras Red Sheep, subcutaneous and omental AD-MSCs took six days to reach same percentage in P1. The initial culture contained heterogeneous cell population with round and fibroblastic cell, upon culture, the number of rounded cell decreased and fibroblastic cells started increasing.…”

“…According to Beaulah et al, (2016) and Fadel et al, (2011), the plastic adherence of AD-MSC was observed after 24 hours in primary culture. Similarly in buffalo, adherence was noticed after 24 hours (Hepsibha et al, 2011).…”

“…Viable AD-MSCs from resuspended pellet were seeded at a density of 1x10 6 cells per T25 culture flask and incubated in 5ml of DMEM-HG at 37 O C under 5 per cent CO 2 in CO 2 incubator (Beaulah et al, 2016) (Figure 4). The spent medium was replaced initially with fresh DMEM-HG after two days of seeding.…”

Comparing the stemness and morphobiometry of spermatogonial stem cells from Doom pig on different days of culture

“…Hoechst or propidium iodide was added and kept at room temperature for 1 min for counterstaining. After washing three times, the plate was examined under a fluorescence microscope (Beaulah et al 2016). Negative controls without primary antibody were used to determine if the secondary antibody is binding nonspecifically to cellular components, resulting in false positives or nonspecific binding.…”